Evaluation of sample preparation protocols for spider venom
profiling by MALDI-TOF MS

J 2017

Evaluation of sample preparation protocols for spider venom profiling by MALDI-TOF MS

BOČÁNEK, Ondřej, Ondrej ŠEDO, Stanislav PEKÁR and Zbyněk ZDRÁHAL

Basic information

Original name

Evaluation of sample preparation protocols for spider venom profiling by MALDI-TOF MS

Authors

BOČÁNEK, Ondřej (203 Czech Republic, belonging to the institution), Ondrej ŠEDO (203 Czech Republic, belonging to the institution), Stanislav PEKÁR (703 Slovakia, guarantor, belonging to the institution) and Zbyněk ZDRÁHAL (203 Czech Republic, belonging to the institution)

Edition

Toxicon, Oxford, England, PERGAMON-ELSEVIER SCIENCE LTD, 2017, 0041-0101

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10406 Analytical chemistry

Country of publisher

United Kingdom of Great Britain and Northern Ireland

Confidentiality degree

není předmětem státního či obchodního tajemství

Impact factor

Impact factor: 2.352

RIV identification code

RIV/00216224:14310/17:00094825

Organization unit

Faculty of Science

DOI

http://dx.doi.org/10.1016/j.toxicon.2017.04.008

UT WoS

000403864000003

Keywords in English

Venom mass fingerprinting; Spider venom; MALDI-TOF MS

Abstract

V originále

Spider venoms are highly complex mixtures containing biologically active substances with potential for use in biotechnology or pharmacology. Fingerprinting of venoms by Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) is a thriving technology, enabling the rapid detection of peptide/protein components that can provide comparative information. In this study, we evaluated the effects of sample preparation procedures on MALDI-TOF mass spectral quality to establish a protocol providing the most reliable analytical outputs. We adopted initial sample preparation conditions from studies already published in this field. Three different MALDI matrixes, three matrix solvents, two sample deposition methods, and different acid concentrations were tested. As a model sample, venom from Brachypelma albopilosa was used. The mass spectra were evaluated on the basis of absolute and relative signal intensities, and signal resolution. By conducting three series of analyses at three weekly intervals, the reproducibility of the mass spectra were assessed as a crucial factor in the selection for optimum conditions. A sample preparation protocol based on the use of an HCCA matrix dissolved in 50% acetonitrile with 2.5% TFA deposited onto the target by the dried-droplet method was found to provide the best results in terms of information yield and repeatability. We propose that this protocol should be followed as a standard procedure, enabling the comparative assessment of MALDI-TOF MS spider venom fingerprints.

Links

GA15-14762S, research and development project

Name: Koevoluce potravní-specializace a složení jedu u pavouků

Investor: Czech Science Foundation

LM2015043, research and development project

Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

Investor: Ministry of Education, Youth and Sports of the CR

LQ1601, research and development project

Name: CEITEC 2020 (Acronym: CEITEC2020)

Investor: Ministry of Education, Youth and Sports of the CR

Citovat

BOČÁNEK, Ondřej, Ondrej ŠEDO, Stanislav PEKÁR and Zbyněk ZDRÁHAL. Evaluation of sample preparation protocols for spider venom profiling by MALDI-TOF MS. Toxicon. Oxford, England: PERGAMON-ELSEVIER SCIENCE LTD, 2017, vol. 133, July, p. 18-25. ISSN 0041-0101. Available from: https://dx.doi.org/10.1016/j.toxicon.2017.04.008.

@article{1384857,
   author = {Bočánek, Ondřej and Šedo, Ondrej and Pekár, Stanislav and Zdráhal, Zbyněk},
   article_location = {Oxford, England},
   article_number = {July},
   doi = {http://dx.doi.org/10.1016/j.toxicon.2017.04.008},
   keywords = {Venom mass fingerprinting; Spider venom; MALDI-TOF MS},
   language = {eng},
   issn = {0041-0101},
   journal = {Toxicon},
   title = {Evaluation of sample preparation protocols for spider venom profiling by MALDI-TOF MS},
   volume = {133},
   year = {2017}
}

TY  - JOUR
ID  - 1384857
AU  - Bočánek, Ondřej - Šedo, Ondrej - Pekár, Stanislav - Zdráhal, Zbyněk
PY  - 2017
TI  - Evaluation of sample preparation protocols for spider venom profiling by MALDI-TOF MS
JF  - Toxicon
VL  - 133
IS  - July
SP  - 18-25
EP  - 18-25
PB  - PERGAMON-ELSEVIER SCIENCE LTD
SN  - 00410101
KW  - Venom mass fingerprinting
KW  - Spider venom
KW  - MALDI-TOF MS
N2  - Spider venoms are highly complex mixtures containing biologically active substances with potential for use in biotechnology or pharmacology. Fingerprinting of venoms by Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) is a thriving technology, enabling the rapid detection of peptide/protein components that can provide comparative information. In this study, we evaluated the effects of sample preparation procedures on MALDI-TOF mass spectral quality to establish a protocol providing the most reliable analytical outputs. We adopted initial sample preparation conditions from studies already published in this field. Three different MALDI matrixes, three matrix solvents, two sample deposition methods, and different acid concentrations were tested. As a model sample, venom from Brachypelma albopilosa was used. The mass spectra were evaluated on the basis of absolute and relative signal intensities, and signal resolution. By conducting three series of analyses at three weekly intervals, the reproducibility of the mass spectra were assessed as a crucial factor in the selection for optimum conditions. A sample preparation protocol based on the use of an HCCA matrix dissolved in 50% acetonitrile with 2.5% TFA deposited onto the target by the dried-droplet method was found to provide the best results in terms of information yield and repeatability. We propose that this protocol should be followed as a standard procedure, enabling the comparative assessment of MALDI-TOF MS spider venom fingerprints.
ER  -

BOČÁNEK, Ondřej, Ondrej ŠEDO, Stanislav PEKÁR and Zbyněk ZDRÁHAL. Evaluation of sample preparation protocols for spider venom profiling by MALDI-TOF MS. \textit{Toxicon}. Oxford, England: PERGAMON-ELSEVIER SCIENCE LTD, 2017, vol.~133, July, p.~18-25. ISSN~0041-0101. Available from: https://dx.doi.org/10.1016/j.toxicon.2017.04.008.

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